Research ArticleMarket Assessment and Product Evaluation ofProbiotic Containing Dietary Supplements Available inBangladesh Market

Anjuman Ara Begum,1 D. M. Jakaria,1 Sharif Md Anisuzzaman,1

Mahfuzul Islam,1 and Siraje Arif Mahmud2

1Department of Pharmacy, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh2Department of Biotechnology and Genetic Engineering, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh

Correspondence should be addressed to Anjuman Ara Begum; anjumantanya@yahoo.com

Received 11 July 2015; Accepted 25 October 2015

Academic Editor: Andrea Masotti

Copyright © 2015 Anjuman Ara Begum et al. This is an open access article distributed under the Creative Commons AttributionLicense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properlycited.

Probiotics containing food supplements available in Bangladesh market were identified and collected for assessment. To assesstheir label claim, they were resuspended into sterile distilled water. Then, series dilutions of each sample solution were preparedand immediately plated out, in duplicate, into De Man Rogosa Sharpe (MRS) agar. These plates were then incubated at 37∘C for 48hours and colonies were counted. Viable cell numbers stated on the labels were compared with actual viable cell numbers. To assessthe viability of the probiotics included in the products, probiotic strains were isolated from each of the four products and screenedfor inhibitory activity against six indicator strains. It was surprisingly found that although the viable cell numbers of all supplementswere three to four log cycles lower than label claim of the products, however, this problem did not affect the inhibitory activity of theprobiotic strains against indicator strains according to in vitro assessment. Legislation and regulation regarding prebiotic-probioticcontaining products should be built up in Bangladesh to ensure quality products supply to the consumers.Moreover, manufacturersof probiotic containing products should take the responsibility for providing the consumer with scientifically and legally correctinformation.

1. Introduction

The role of the diet is to provide nutrients to meet host phys-iological requirements. The concept of functional foods hasbeen evolved recently after extensive research to find out therole of nutrients on health [1]. Functional foods contain phys-iologically active components, which provide health benefitsbeyond basic nutrition by affecting one or more functions inthe body in a targeted way [2]. “The functional componentcould include essential macronutrient with specific physio-logical effect and components that have some nutritive valuebut are not classified as ‘essential’, such as oligosaccharides,or food components with no nutritive value, such as livemicroorganisms or plant chemicals” [3]. A dietary supple-ment is a product intended for ingestion as a supplement to

the diet and it can be manufactured as pills, tablets, capsules,gel caps, liquids, and powders [4]. Dietary supplements orfunctional foods contain vitamins, micronutrients, antioxi-dants, certain bioactive peptides, herbs, polyunsaturated fattyacids, and so forth. The concept has now moved markedlytowards gastrointestinal function, in particular the impactof gut bacteria. The most frequently used dietary method ofinfluencing the gut flora composition is that of probiotics.Probiotics are live microorganisms (in most cases, bacteria)that are similar to beneficial microorganisms found in thehuman gut. According to FAO/WHO (Food and AgricultureOrganization andWorldHealthOrganization), probiotics aredefined as “live microorganisms which when administered inadequate amounts confer a health benefit on the host” [5].Over 100,000 billion bacteria (more than 500 species) live in

Hindawi Publishing CorporationJournal of PharmaceuticsVolume 2015, Article ID 763796, 5 pageshttp://dx.doi.org/10.1155/2015/763796

2 Journal of Pharmaceutics

the human gut. Human gut flora contains friendly and harm-ful bacteria. The metabolism products of friendly bacteria(probiotic) such as lactic acid and acetic acid can inhibit thegrowth of harmful bacteria and confer health benefits on thehost [6]. Probiotics are ingested for their purported positiveadvantages in the digestive tract and/or systemic areas like theliver, vagina, or bloodstream, for example, neutralization oftoxins, increase of the immune response [7, 8], antimutagenicand anticarcinogenic activities [9–11], reduction of choles-terol levels [12], control of diarrhea [13], alleviation of lactoseintolerance [14], and inflammatory bowel diseases [15]. Theyare also a source of vitamins, especially of the B group [16,17], and can also be used as complementary and alternativemedicine (CAM) [18]. Consumers should be provided withan independent assessment of physiological, microbial, andsafety aspects of these live microbial products, especially ifthey can improve health. The latest trend in the functionalfood market is to combine probiotics with prebiotics toenhance the effect of probiotics [19]. Prebiotics are definedas “nondigestible food ingredients that beneficially affect thehost by selectively stimulating the growth and/or activity ofprobiotic bacteria in the colon” [1]. Probiotic trials shoulduse the best methodologies available. For probiotics to exertbeneficial properties, they must have a high viability in theproduct and have robust survival properties in the gut, whichis their first point of contact. Because of the presence ofpotentially pathogenic species such as Enterococcus faeciumand E. faecalis in probiotic products [2, 19], the productionand marketing of functional foods should be strictly con-trolled and carefullymonitored [20]. Information on the labelof the product, especially regarding the composition andidentity of the probiotic strains included, needs to be accurateto guarantee safety and functionality [21]. In most cases,the identity and number of viable strains recovered did notcorrespond with the information on the label. In view of theabove, the aims of this study were to complete a market andproduct assessment of probiotic containing food supplementsavailable in Bangladesh.This involved identifying the range ofproducts available on the Bangladesh market and evaluatingclaims made on the labels.

2. Methodology

2.1. Collection of Sample. To conduct the study, different phar-macies, super shops, and medicine corners were surveyedto find out probiotic containing dietary supplements. Onlyfour different probiotic containing dietary supplements wereidentified and collected.

2.2. Microbial Assessment of Viable Cell Numbers Included inFour Selected Probiotic Supplements. Four probiotic sup-plements readily available in pharmacies were selected todetermine the viability (growth) of the probiotic strains andto compare actual viable cell numbers with the label claim inthis regard. The content of the capsule was resuspended in10mL sterile distilled water. After performing serial dilution,appropriately diluted solutions were than plated out, induplicate, onto a De Man Rogosa Sharp (MRS) agar. Platesfrom each dilution were incubated at 37∘C and colonies werecounted after 48 hours.

2.3. Assessment of the Viability of the Probiotics Available inthe Products. To assess the viability of the probiotics includedin the products, probiotic strains isolated from each of the4 products were screened for inhibitory activity against thefollowing 6 indicator strains: Salmonella typhi, Salmonellasp., Shigella sp., Staphylococcus aureus, Escherichia coli, andVibrio cholerae. The probiotic strains were cultured in MRbroth (Biolab) for 18 hours at 37∘C and 10 𝜇L was spotted onMRS agar (Biolab). The plates were incubated for 24 hoursat 37∘C and then lawned with active cells of the indicatorstrains (approximately 106 cfu/mL), embedded in soft agar(0.8%, m/v). The plates were incubated at 37∘C for 24 hoursand the colonies were examined for the formation of zones,which indicates the level of inhibitory activity and thereforeviability. The study was done in triplicate and the averageswere determined.

3. Results and Discussions

The prebiotic and probiotic containing product market isa fast-growing industry worldwide and the list of avail-able products increases on a daily basis. Although as adeveloping country, Bangladesh has limited scope and facil-ity but is trying to cope up with the health demand ofprobiotic. One of the leading pharmaceutical companies hasrealized the importance of functional food and launched aprobiotics product in capsule form. Throughout the worldprebiotics and probiotics are available in fermented foodlike yoghurt and also in baby foods. Through the marketassessment, different types of food products were foundcontaining probiotics and combination of probiotics andprebiotics. Among them some are fortified baby food, dairyproducts, and yoghurt products. But only four food supple-ments were found in Bangladesh containing only probiotics.Among them one product is manufactured in Bangladeshand two capsules were from India and one sachet (powder)was from Taiwan. Three samples were of capsule form andonly one was in powdered form available in sachet. Butno food supplements were found having probiotic and pre-biotic combination. All the four samples contained sameprobiotics like textit Lactobacillus acidophilus, Lactobacillusbulgaricus, Bifidobacterium bifidum, Lactobacillus salivarius,Lactobacillus casei, Bifidobacterium longum, Saccharomycesboulardii, Lactobacillus rhamnosus, Bifidobacterium infan-tis, Lactococcus lactis, Lactobacillus paracasei, and Streptococ-cus thermophilus but the viable cell numbers are different.As a functional food component, prebiotics and probioticsare conceptually intermediate between foods and drugs.Depending on the jurisdiction, they typically receive anintermediate level of regulatory scrutiny, in particular ofthe health claims made concerning them. For probiotic toexert beneficial properties, they must have a high viabilityin the product and have robust survival properties in thegut, which is their first point of contact [22]. That is whyassessment of the label claim is necessary for these foodproducts. Comparison of viable cell numbers stated on thelabels of the supplements with the actual viable cell numbersis presented in Table 1 and Figure 1.

Journal of Pharmaceutics 3

(Supplement 1) (Supplement 2)

(Supplement 3) (Supplement 4)

(Supplement 1) (Supplement 2)

Figure 1: A partial presentation of growth of probiotics of four different products.

Table 1: Comparison of the actual viable cell numbers with theclaims on the tables of 4 probiotic supplements available in Bang-ladesh market.

Supplement

Viable cell numbersstated on the labelof the supplement

(cfu/g)

Actual viable cellnumbers identified (cfu/g)

1 4 × 109 3.98 × 106 ± 0.03∗

2 5 × 109 3.19 × 105 ± 0.5∗

3 2.5 × 109 1.96 × 106 ± 0.04∗

4 1 × 1010 1 × 107 ± 0.01∗∗Indicates values that are significantly lower than the corresponding viablecell numbers stated on the label of the supplement (𝑃 < 0.05 after indepen-dent sample 𝑡-test).

To achieve health benefits, probiotic bacteria must beviable and available at a high concentration, typically 106–107 cfu/g of product [23]. From this investigation, it was found

that viable cell numbers of all supplements were three tofour log cycles lower than the claimed number and threeof the four products are marginally potential for their pro-posed health benefit purpose. In different countries, despitethe importance of viability, surveys conducted to validateviability claims have shown low populations of probioticbacteria in probiotic products [24]. Several factors have beenclaimed to be responsible for the loss of viability of probi-otic organisms: acidity of products, acid produced duringrefrigerated storage (after acidification), level of oxygen inproducts, oxygen permeation through the package, andsensitivity to antimicrobial substances produced by bacteria[25].

To determine the inhibitory activity of probiotics, sixindicator bacterial strains are used.They are Salmonella typhi,Salmonella sp., Shigella sp., Staphylococcus aureus, Escherichiacoli, and Vibrio cholerae. The result of the screening of theprobiotic strains isolated from the 4 selected supplementsagainst 6 indicator strains is presented in Table 2. It is evidentthat all strains showed good inhibitory activity.

4 Journal of Pharmaceutics

Table 2: Inhibitory activity of probiotic strains from 4 probiotic supplements against a panel of indicator strains.

Human pathogen Zone size (mm)

Supplement 1 Supplement 2 Supplement 3 Supplement 4

Salmonella typhi 26–30 26–31 26–32 26–30

Salmonella sp. 31–35 31–33 31-32 31–35

Shigella sp. 22–24 22–25 22-23 22–24

Staphylococcus aureus 18–22 18–20 18–21 18–22

Escherichia coli 24–30 24–30 24–28 24–26

Vibrio cholerae 10–14 10-11 10–14 10–13

Table 2 reveals that the viable probiotic cells showed goodinhibitory activity despite their numbers were lower thanrespective label claims. Therefore, in order to find out theiractual efficacy, in vivo assessment should be considered. Thissituation indicates that as there is no regulation in Bangladeshfor these types of products, the consumers are being manip-ulated by the manufacturers and marketers into buying aproduct under false claim. It is important that the healthclaims stated on the labels of products supply the consumerwith reliable information because such claims influenceconsumer behavior and potentially affect public health [26].Proper manufacturing practice and correct storage should bemaintained by themanufacturers andmarketers to ensure cellsurvival.

4. Conclusion

Different probiotics containing food supplements and vari-eties of fortified baby foods (containing probiotics andprebiotics) are imported in Bangladesh. But in Bangladeshthere is no regulation to assess the quality of functional foodcontaining products.That is why marketers of these productsaremisleading consumerswith a number of health claims thatare not scientifically sound. Now it becomes a prime concernto produce proper legislation and regulation in the field ofprobiotics and prebiotics containing food supplement andalso it should be revised properly, so that quality products ofprebiotic-probiotic or functional food can be manufacturedand marketed for the benefit of consumers.

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper.

Acknowledgments

The authors are thankful to the University Grants Commis-sion of Bangladesh for their financial support and to theDepartment of Pharmacy and theDepartment of Biotechnol-ogy and Genetic Engineering for providing their laboratorysupport to perform the study properly.

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